• Title/Summary/Keyword: Sb addition

Search Result 283, Processing Time 0.033 seconds

The EST Analysis and Transgene Expression System in Rice

  • Kim, Jukon;Nahm, Baek-Hie
    • Journal of Plant Biotechnology
    • /
    • v.1 no.1
    • /
    • pp.46-55
    • /
    • 1999
  • The expressed sequence tags(ESTs) from immature seed of rice, Oryza sativa cv Milyang 23, were partially sequenced and analyzed by homology. As of 1998, the partial sequences of about 6,600 cDNA clones were analyzed from normal and normalized immature seed cDNA libraries. About 2,200 ESTs were putatively identified by BLASTX deduced amino acid sequence homology analysis. About 20% of them were putatively identified as storage proteins. Also the clones were highly homologous to genes involved particularly in starch biosynthesis, glycolysis, signal transduction and defenses. Compared to 35% of redundancy in the ESTs of normal cDNA library, that from the substracted library was 15%. The Korea Rice Genome Network is maintained to provide the updated information of sequences, their homologies and sequence alignments of ESTs. For the stable expression of transgene in rice, diverse vectors were developed for overexpression, targeting and gene dosage effect with transit peptides (Tp) and matrix attachment region (MAR) sequence from chicken lysozyme locus. The rice calli were transformed via Agrobacterium tumefaciens LBA4404(pSB1) with the triparental mating technique and selected by herbicide resistance. The green fluorescent protein(GFP) gene in expression vector under the control of rbcS promoter-Tp was overexpressed upto 10 % of the total soluble protein. In addition, the Tp-sGFP fusion protein was properly processed during translocation into chloroplast. The expression of sGFP in the presence of MAR sequences was analyzed with Northern and immunoblot analysis. All the lines in which sGFP transgene with MAR sequence, showed position independent and copy number-dependent expression, while the lines without MAR showed the varied level of expression with the integration site. Thus the MAR sequence significantly reduced the variation in transgene expression between independent transformants.

  • PDF

Fucosyltransferase IV Enhances Expression of MMP-12 Stimulated by EGF via the ERK1/2, p38 and NF-kB Pathways in A431Cells

  • Yang, Xue-Song;Liu, Shui-Ai;Liu, Ji-Wei;Yan, Qiu
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.4
    • /
    • pp.1657-1662
    • /
    • 2012
  • Fucosyltransferase IV (FUT4) has been implicated in cell adhesion, motility, and tumor progression in human epidermoid carcinoma A431 cells. We previously reported that it promotes cell proliferation through the ERK/MAPK and PI3K/Akt signaling pathways; however, the molecular mechanisms underlying FUT4-induced cell invasion remain unknown. In this study we determined the effect of FUT4 on expression of matrix metalloproteinase (MMP)-12 induced by EGF in A431 cells. Treatment with EGF resulted in an alteration of cell morphology and induced an increase in the expression of MMP-12. EGF induced nuclear translocation of nuclear factor kB (NF-${\kappa}B$) and resulted in phosphorylation of $IkB{\alpha}$ in a time-dependent manner. In addition, ERK1/2 and p38 MAPK were shown to play a crucial role in mediating EGF-induced NF-${\kappa}B$ translocation and phosphorylation of $I{\kappa}B{\alpha}$ when treated with the MAPK inhibitors, PD98059 and SB203580, which resulted in increased MMP-12 expression. Importantly, we showed that FUT4 up-regulated EGF-induced MMP-12 expression by promoting the phosphorylation of ERK1/2 and p38 MAPK, thereby inducing phosphorylation/degradation of $I{\kappa}B{\alpha}$, NF-${\kappa}B$ activation. Base on our data, we propose that FUT4 up-regulates expression of MMP-12 via a MAPK-NF-${\kappa}B$-dependent mechanism.

Anti-Proliferative Activity of Ethanol Extracts from Taxilli Ramulus (Taxillus chinensis (DC.) Danser) Through Cyclin D1 Proteasomal Degradation in Human Colorectal Cancer Cells

  • Park, Gwang Hun;Song, Hun Min;Park, Su Bin;Park, Ji Hye;Shin, Myeong Su;Son, Ho-Jun;Um, Yurry;Jeong, Jin Boo
    • Korean Journal of Plant Resources
    • /
    • v.30 no.6
    • /
    • pp.640-646
    • /
    • 2017
  • In this study, we elucidated anti-cancer activity and potential molecular mechanism of 70% ethanol extracts from Taxilli Ramulus (Taxillus chinensis (DC.) Danser) (TR-E70) against human colorectal cancer cells. Anti-cell proliferative effect of TR-E70 was evaluated by MTT assay. The effect of TR-E70 on the expression of cyclin D1 in the protein and mRNA level was evaluated by Western blot and RT-PCR, respectively. TR-E70 suppressed the proliferation of human colorectal cancer cell lines, HCT116 and SW480. Although TR-E70 decreased cyclin D1 expression in protein and mRNA level, decreased level of cyclin D1 protein by TR-E70 more dramatically occurred than that of cyclin D1 mRNA. Cyclin D1 downregulation by TR-E70 was attenuated in presence of MG132. In addition, TR-E70 phosphorylated threonine-286 (T286) of cyclin D1. TR-E70-mediated cyclin D1 degradation was blocked in presence of LiCl as an inhibitor $GSK3{\beta}$ but not PD98059 as an ERK1/2 inhibitor and SB203580 as a p38 inhibitor. Our results suggest that TR-E70 may downregulate cyclin D1 as one of the potential anti-cancer targets through $GSK3{\beta}$-dependent cyclin D1 degradation. From these findings, TR-E70 has potential to be a candidate for the development of chemoprevention or therapeutic agents for human colorectal cancer.

Protective Effects of Samul-tang on ${H_2O_2}-induced$ Cell Apoptosis in Cultured Cardiomyoblast Cells ($H_2O_2$에 의한 배양심근세포고사에 미치는 사물탕의 방어효과)

  • 박종운;한상혁;김도환;문병순
    • The Journal of Korean Medicine
    • /
    • v.22 no.4
    • /
    • pp.58-68
    • /
    • 2001
  • Objectives : This study was designed to investigate the protective mechanisms of Samul-tang (SMT) on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Methods : The cultured cells were pretreated with SMT and exposed to $H_2O_2$. The cell damage was assessed by using MTT assay. Also, we used Hoechst staining, Western blotting analysis. Results : SMT significantly reduced both $H_2O_2$-induced cell death and chromatin fragmentation. The decrease of Bcl2 expression by $H_2O_2$ was inhibited by SMT. In addition, the increase of Bax expression was also inhibited by SMT. In particular, Fas expression, which is generally recognized as cell death inducing signal by Fas/FasL interaction, was markedly decreased by $H_2O_2$ in a time-dependent manner, whereas this decrease was completely prevented by SMT. The cotreatment of SMT and $H_2O_2$ in H9c2 cells also induced the phosphorylation of ERK in a time-dependent manner. Moreover, PD098059, a specific inhibitor of ERKl/2, attenuated the protective effect of SMT on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Furthermore, the protective effect of SMT was significantly blocked by treatment of SB203580, a specific inhibitor of p38. Conclusions : Taken together, this study suggests that the protective effects of the water extract of SMT against oxidative damages may be mediated by the modulation of Bel2 and Bax expression via the regulation of ERK and p38 signaling pathway.

  • PDF

The Cholesterol-Binding Antibiotic Nystatin Induces Expression of Macrophage Inflammatory Protein-1 in Macrophages

  • Baek, Seungil;Kim, Sun-Mi;Lee, Sae-A;Rhim, Byung-Yong;Eo, Seong-Kug;Kim, Koanhoi
    • Biomolecules & Therapeutics
    • /
    • v.21 no.1
    • /
    • pp.42-48
    • /
    • 2013
  • Nystatin, a polyene antifungal antibiotic, is a cholesterol sequestering agent. The antifungal agent alters composition of the plasma membrane of eukaryotic cells, whereas its effects on cells are poorly investigated. In the current study, we investigated the question of whether nystatin was able to induce expression of macrophage inflammatory protein-1 (MIP-1). THP-1 cells rarely express MIP-$1{\alpha}$ and MIP-$1{\beta}$, however, upon exposure to nystatin, significantly elevated expression of MIP-$1{\alpha}$ and MIP-$1{\beta}$ was observed in a dose-dependent fashion at the messenger and protein levels. Cellular factors activated by nystatin as well as involved in nystatin-induced expression of MIP-1 proteins were identified in order to understand the molecular mechanisms of action of the anti-fungal agent. Treatment with nystatin resulted in enhanced phosphorylation of Akt, ERK, p38 MAPK, and JNK. Abrogation or significant attenuation of nystatin-induced expression of MIP-$1{\alpha}$ and MIP-$1{\beta}$ was observed by treatment with Akt inhibitor IV, LY294002, and SP6001250. Inhibition of ERK or p38MAPK using U0126 and SB202190 did not lead to attenuation of MIP-1 expression. In addition, inhibitors of protein kinase C, such as GF109203X and Ro-318220, also attenuated expression of MIP-1. These results indicate that nystatin is able to activate multiple cellular kinases and, among them, Akt and JNK play primary roles in nystatin-induced expression of MIP-1 proteins.

Enhancing Effect of Extracts of Phellodendri Cortex on Glucose Uptake in Normal and Insulin-resistant 3T3-L1 Adipocytes (3T3-L1 지방세포에서 황백 추출물의 Glucose Uptake 촉진 및 인슐린 저항성 개선 활성)

  • Kim, So-Hui;Shin, Eun-Jung;Hyun, Chang-Kee
    • Korean Journal of Pharmacognosy
    • /
    • v.36 no.4 s.143
    • /
    • pp.291-298
    • /
    • 2005
  • Anti-hyperglycemic effects of 17 medicinal plants that have been used for ameliorating diabetes in oriental medicine were evaluated using glucose transport assay in 3T3-L1 adipocytes. Higher activities were obtained by treating water or alcohol extract of Phellodendri Cortex (PC), which showed enhancing effects both on basal and insulin-stimulated glucose uptake. The latter effect of PC was completely inhibited by wortmannin, a specific inhibitor for phosphatidyl inositol 3-kinase (PI 3-kinase), but not affected by SB203580, A specific inhibitor for p38 mitogen-activatedprotein kinase(MAPK). Genistein, an inhibitor for tyrosine kinases, abolished the PC effects completely. Treatment of vanadate, an inhibitor for tyrosine phosphatases, together with PC showed no significant synergic enhancement in glucose uptake. The results of inhibitors associated with insulin signaling pathway indicated that extracts of PC enhance glucose uptake by PI-3 kinase activation which is an upstream event for GLUT4 translocation. Antidiabetic effects of PC extract might be also due to enhanced tyrosine phosphorylation and reduced tyrosine dephosphorylation. In addition, PC accelerated insulin-stimulated glucose uptake in insulin-resistant cells, recovering the uptake level close to that of normal cells. These findings may offer a new way to utilize extracts of PC as novel anti-hyperglycemic agents.

Piezoelectric and Dielectric Properties on PSN-PMN-PZT Composition according to CeO2 Addition (PSN-PMN-PZT 조성의 CeO2첨가에 따른 압전.유전특성 변화)

  • Yoon, Man-Soon;Chio, Yong-Gil;Ur, Soon-Chul
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
    • /
    • v.19 no.9
    • /
    • pp.838-842
    • /
    • 2006
  • 0.03Pb$(Sb_{0.5}Nb_{0.5})O_{3}-0.03Pb(Mn{1/3}Nb{2/3)O_{3}-(0.94-x)PbTiO_{3}-xPbZrO_{3}$ ceramics doped with $CeO_{2}$ were synthesized by conventional bulk ceramic processing technique. Phases analysis, microstructures and piezoelectric properties were investigated as a function of $CeO_{2}$ content (0.03, 0.05, 0.1 0.3, 0.5 and 0.7 wt%). Microstructures and phases information were characterized using a scanning electron microscope (SEM) and an X-ray diffractometer (XRD). Mechanical quality factor ($Q_{m}$) and coupling factor(kp) were obtained from the resonance measurement method. Both $Q_{m}$ and $k_{p}$ were shown to reach to the maximum at 0.1 wt% $CeO_{2}$. In order to evaluate the stability of resonance frequency and effective electromechanical coupling factor ($K_{eff}$) as a function of $CeO_{2}$, the variation of resonance and anti-resonance frequency were also measured using a high voltage frequency response analyzer under various alternating electric fields from 10 V/mm to 80 V/mm. It was shown that the stability of resonance frequency and effective electromechanical coupling factor were increased with increasing the $CeO_{2}$ contents.

High Density Inductively Coupled Plasma Etching of III-V Semiconductors in BCI3Ne Chemistry (BCI3Ne 혼합가스를 이용한 III-V 반도체의 고밀도 유도결합 플라즈마 식각)

  • 백인규;임완태;이제원;조관식
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
    • /
    • v.16 no.12S
    • /
    • pp.1187-1194
    • /
    • 2003
  • A BCl$_3$/Ne plasma chemistry was used to etch Ga-based (GaAs, AIGaAs, GaSb) and In-based (InGaP, InP, InAs and InGaAsP) compound semiconductors in a Planar Inductively Coupled Plasma (ICP) reactor. The addition of the Ne instead of Ar can minimize electrical and optical damage during dry etching of III-V semiconductors due to its light mass compared to that of Ar All of the materials exhibited a maximum etch rate at BCl$_3$ to Ne ratios of 0.25-0.5. Under all conditions, the Ga-based materials etched at significantly higher rates than the In-based materials, due to relatively high volatilities of their trichloride etch products (boiling point CaCl$_3$ : 201 $^{\circ}C$, AsCl$_3$ : 130 $^{\circ}C$, PCl$_3$: 76 $^{\circ}C$) compared to InCl$_3$ (boiling point : 600 $^{\circ}C$). We obtained low root-mean-square(RMS) roughness of the etched sulfate of both AIGaAs and GaAs, which is quite comparable to the unetched control samples. Excellent etch anisotropy ( > 85$^{\circ}$) of the GaAs and AIGaAs in our PICP BCl$_3$/Ne etching relies on some degree of sidewall passivation by redeposition of etch products and photoresist from the mask. However, the surfaces of In-based materials are somewhat degraded during the BCl$_3$/Ne etching due to the low volatility of InCl$_{x}$./.

Preliminary study on car detection and tracking method using surveillance camera in tunnel environment for accident detection (터널 내 유고상황 자동 판정을 위한 선행 연구: CCTV를 이용한 차량의 탐지와 추적 기법 고찰)

  • Oh, Young-Sup;Shin, Hyu-Soung
    • Journal of Korean Tunnelling and Underground Space Association
    • /
    • v.19 no.5
    • /
    • pp.813-827
    • /
    • 2017
  • Surveillance cameras installed in tunnels capture the various video frames effected by dynamic and variable factors. In addition, localizing and managing the cameras in tunnel is not affordable, and quality of capturing frame is effected by time. In this paper, we introduce a new method to detect and track the vehicles in tunnel by using surveillance cameras installed in a tunnel. It is difficult to detect the video frames directly from surveillance cameras due to the motion blur effect and blurring effect on lens by dirt. In order to overcome this difficulties, two new methods such as Differential Frame/Non-Maxima Suppression (DFNMS) and Haar Cascade Detector to track cars are proposed and investigated for their feasibilities. In the study, it was shown that high precision and recall values could be achieved by the two methods, which then be capable of providing practical data and key information to an automatic accident detection system in tunnels.

Nerve Growth Factor Activates Brain-derived Neurotrophic Factor Promoter IV via Extracellular Signal-regulated Protein Kinase 1/2 in PC12 Cells

  • Park, So Yun;Lee, Ji Yun;Choi, Jun Young;Park, Mae Ja;Kim, Dong Sun
    • Molecules and Cells
    • /
    • v.21 no.2
    • /
    • pp.237-243
    • /
    • 2006
  • Brain-derived neurotrophic factor (BDNF) is a neuromodulator of nociceptive responses in the dorsal root ganglia (DRG) and spinal cord. BDNF synthesis increases in response to nerve growth factor (NGF) in trkA-expressing small and medium-sized DRG neurons after inflammation. Previously we demonstrated differential activation of multiple BDNF promoters in the DRG following peripheral nerve injury and inflammation. Using reporter constructs containing individual promoter regions, we investigated the effect of NGF on the multiple BDNF promoters, and the signaling pathway by which NGF activates these promoters in PC12 cells. Although all the promoters were activated 2.4-7.1-fold by NGF treatment, promoter IV gave the greatest induction. The p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, phosphatidylinositol 3-kinase (PI-3K) inhibitor, LY294003, protein kinase A (PKA) inhibitor, H89, and protein kinase C (PKC) inhibitor, chelerythrine, had no effect on activation of promoter IV by NGF. However, activation was completely abolished by the MAPK kinase (MEK) inhibitors, U0126 and PD98059. In addition, these inhibitors blocked NGF-induced phosphorylation of extracellular signal-regulated protein kinase (ERK) 1/2. Taken together, these results suggest that the ERK1/2 pathway activates BDNF promoter IV in response to NGF independently of NGF-activated signaling pathways involving PKA and PKC.